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Nanomanipulation of analysis of lipoplex assembly

脂质复合物组装分析的纳米操作

基本信息

批准号：
7886754
负责人：
VALENTIN V RYBENKOV
金额：
$ 18.56万
依托单位：
UNIVERSITY OF OKLAHOMA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-06 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7886754
关键词：
Address Adverse effects Affect Age Animal Model Cells Charge Collaborations Complex DNA DNA delivery Data Development Diabetes Mellitus Disease Drug Formulations Ensure Fatty Acids Future Goals Head Heart Hereditary Disease Inherited Intervention Learning Lipid Biochemistry Lipids Lysosomal Storage Diseases Malignant Neoplasms Mammalian Cell Membrane Lipids Methods Nucleic Acids Physical Chemistry Physical condensation Process Property Research Research Personnel Retinal Degeneration Series Solutions Superhelical DNA System Tail Techniques Testing Transfection Vesicle Viral Virus base cell assembly clinical application clinically relevant design gene therapy improved nanoparticle nervous system disorder neuroinflammation novel pre-clinical public health relevance research study single molecule skills success targeted delivery tool vector viral DNA

项目摘要

DESCRIPTION (provided by applicant): Gene therapy holds a promise to treat a variety of genetic disorders including cancer, diabetes and many other inherited and acquired diseases. Successful clinical application of gene therapy requires the development of efficient tools for targeted delivery of nucleic acids into affected cells. Presently used virus-based DNA delivery systems produce serious immunological and oncological side effects. Lipid-based DNA delivery systems are widely viewed as a potentially safer alternative. The development of effective lipid-based DNA nanoparticles (lipoplexes) is limited by poor understanding of how DNA is packed into the lipid vesicles. This project aims at characterizing the assembly of lipoplexes using single DNA manipulations. Preliminary studies revealed a number of mechanistic details of lipid-DNA interaction that were not detected using more conventional, bulk methods. We plan to evaluate the idea that the interactions of lipids with DNA can be correlated with the efficiency of lipoplex assembly and cell transfection efficacy. Specific aims of the project are (i) Quantify deformation of single DNA by DOTAP, (ii) compare a series of lipids in their ability to compact DNA; (iii) correlate the single molecule data with the quality of assembled lipoplexes. The completion of this study will evaluate the idea that single molecule data can improve the assembly of lipoplexes and transfection efficiency. If true, this would make the assembly of DNA containing nanoparticles a much more predictable process and would lay grounds for more rational design of non-viral DNA delivery systems. This in turn may help develop lipoplexes suitable for clinical applications and thereby enable novel interventions in treating disease. PUBLIC HEALTH RELEVANCE: Gene therapy could potentially be used to treat a variety of inherited and acquired ailments including cancer, diabetes, aging and heart and neurological disorders. The development of non-viral, lipid-based DNA delivery vectors could help design efficient gene therapies with reduced side effects. We plan to evaluate the idea that the design of such vectors can be greatly improved using single DNA nanomanipulation technique.

描述（由申请人提供）：基因疗法有望治疗多种遗传性疾病，包括癌症、糖尿病和许多其他遗传性和获得性疾病。基因治疗的成功临床应用需要开发有效的工具，将核酸靶向递送到受影响的细胞中。目前使用的基于病毒的 DNA 传递系统会产生严重的免疫学和肿瘤学副作用。基于脂质的 DNA 传递系统被广泛视为一种可能更安全的替代方案。由于对 DNA 如何填充到脂质囊泡中的了解不足，有效的基于脂质的 DNA 纳米颗粒（lipoplex）的开发受到限制。该项目旨在利用单一 DNA 操作来表征脂质复合物的组装。初步研究揭示了脂质-DNA 相互作用的许多机制细节，而这些细节是使用更传统的批量方法无法检测到的。我们计划评估脂质与 DNA 的相互作用与脂质复合物组装效率和细胞转染功效相关的想法。该项目的具体目标是 (i) 通过 DOTAP 量化单个 DNA 的变形，(ii) 比较一系列脂质压缩 DNA 的能力； (iii) 将单分子数据与组装的脂质复合物的质量相关联。这项研究的完成将评估单分子数据可以提高脂质复合物的组装和转染效率的想法。如果属实，这将使含有纳米粒子的 DNA 组装成为一个更加可预测的过程，并为更合理地设计非病毒 DNA 传递系统奠定基础。这反过来可能有助于开发适合临床应用的脂质复合物，从而实现治疗疾病的新干预措施。公共健康相关性：基因疗法有可能用于治疗各种遗传性和后天性疾病，包括癌症、糖尿病、衰老以及心脏和神经系统疾病。非病毒、基于脂质的 DNA 递送载体的开发有助于设计有效的基因疗法，并减少副作用。我们计划评估使用单 DNA 纳米操作技术可以大大改进此类载体的设计的想法。